大鼠骨髓内皮祖细胞SPIO标记与检测.pdfVIP

山西医科大学硕士学位论文 Rat bone endothelial progenitor cells labeled with superparamagnetic iron oxide particles and the detection Abstract Obiective To study the rat bone marrow progenitor cells (endothelial progenitor cells, EPCS) the isolation, culture, SPIO mark, and observe whether there is any effect on cellular viability、 proliferation and apoptosis. which lay the foundation for future experimental studies. Methods The EPCS were collected from bone marrow of rats(weighing120g~150g)and cultured. EPCS were obtained by density gradient centrifugation and adhesive-screening methods and ／ labeled with 25μg mL SPIO. Marked and unlabeled groups compared: Prussian blue staining was used to identify labeling index.Trypan blue staining was employed to detect cell vitality. MTT assay was utilized to detect proliferation activity of stem cells; Calcein - AM/PI staining cell was detected cell activity and membrane integrity; AO / PI staining was detected cell apoptosis. Results Cell Colonies are connected, showing the typical cobblestone appearance on the 7 ~ 10 d. Prussian blue staining showed that SPIO (25μg/mL, 24 h) of cells labeled rate of close to 94%; compared SPIO labeled and unlabeled EPCS: cell viability was 94.34% ± 0.25% and 95.33% ± 0.31%; MTT assay detected that there was no statistical difference between the groups compared with Italy in righteousness (P 0.05); Calcein-AM/PI staining cell activity and membrane integrity, survival rates were 96% and 97%; AO / PI staining cells withered groups death rate was 5%. Conclusion Gradient centrifugation from bone marrow mononuclear cells can be achieve